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RecBCD is a model enzyme for the use of single molecule fluorescence as an experimental technique used to better understand the function of protein-DNA interactions. [23] The enzyme is also useful in removing linear DNA, either single- or double-stranded, from preparations of circular double-stranded DNA, since it requires a DNA end for activity.
That requires a second protein, attached to Cas9: a reverse transcriptase enzyme, which can make a new DNA strand from the RNA template and insert it at the nicked site. [269] Those three independent pairing events each provide an opportunity to prevent off-target sequences, which significantly increases targeting flexibility and editing ...
DNA recombinases are widely used in multicellular organisms to manipulate the structure of genomes, and to control gene expression.These enzymes, derived from bacteria (bacteriophages) and fungi, catalyze directionally sensitive DNA exchange reactions between short (30–40 nucleotides) target site sequences that are specific to each recombinase.
In genetics, Flp-FRT recombination is a site-directed recombination technology, increasingly used to manipulate an organism's DNA under controlled conditions in vivo.It is analogous to Cre-lox recombination but involves the recombination of sequences between short flippase recognition target (FRT) sites by the recombinase flippase (Flp) derived from the 2 μ plasmid of baker's yeast ...
RuvA (Holliday junction branch migration complex subunit RuvA) [4] is a DNA-binding protein that binds Holliday junctions with high affinity. The structure of the complex has been variously elucidated through X-ray crystallography and EM data, and suggest that the complex consists of either one or two RuvA tetramers, with charge lined grooves through which the incoming DNA is channelled.
Transformed oligonucleotides are incorporated into the cellular DNA through λ-red- and Cas9 endonuclease-mediated homologous recombination. [ 1 ] λ-red is activated when arabinose binds to expressed AraC , inducing dimerization of the AraC protein and subsequent DNA binding to activate the promoter [ 18 ] prior to oligonucleotide transfection.
Cas9 (or "CRISPR-associated protein 9") is an enzyme that uses CRISPR sequences as a guide to recognize and open up specific strands of DNA that are complementary to the CRISPR sequence. Cas9 enzymes together with CRISPR sequences form the basis of a technology known as CRISPR-Cas9 that can be used to edit genes within living organisms.
[1] [2] [3] Enzymes known as site-specific recombinases (SSRs) perform rearrangements of DNA segments by recognizing and binding to short, specific DNA sequences (sites), at which they cleave the DNA backbone, exchange the two DNA helices involved, and rejoin the DNA strands. In some cases the presence of a recombinase enzyme and the ...